Epitope tags (including small molecules, peptides, and proteins) can be attached to a molecule of interest by using chemical conjugation or genetic fusion. This allows detection of the tagged molecule with antibodies or other specific binding reagents that recognize the tag itself. This makes it easy for tracking protein expression and localization. Epitope tagging also enables affinity purification of the tagged molecule by using tag-specific antibodies coupled to chromatographic resins.
With epitope-tagging, one tag can be coupled to a range of molecules in a series of purifications or detection experiments.
A single anti-tag antibody can then be used to detect the target molecules as an alternative to individual molecule-specific antibodies. This simplifies experimental design and also saves money.
The use of two epitope tags in one experiment can be exploited to evaluate molecular interactions. In this scenario, immunoprecipitation with an antibody against one tag can be followed by Western blot detection with an antibody against the second tag.
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